[go: up one dir, main page]

WO2019105893A1 - Dispositif et procédé d'examen d'une suspension de cellules - Google Patents

Dispositif et procédé d'examen d'une suspension de cellules Download PDF

Info

Publication number
WO2019105893A1
WO2019105893A1 PCT/EP2018/082576 EP2018082576W WO2019105893A1 WO 2019105893 A1 WO2019105893 A1 WO 2019105893A1 EP 2018082576 W EP2018082576 W EP 2018082576W WO 2019105893 A1 WO2019105893 A1 WO 2019105893A1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
type
radiation
affinity molecules
interaction surface
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/EP2018/082576
Other languages
German (de)
English (en)
Inventor
Falk Schlaudraff
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Leica Microsystems CMS GmbH
Original Assignee
Leica Microsystems CMS GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Leica Microsystems CMS GmbH filed Critical Leica Microsystems CMS GmbH
Publication of WO2019105893A1 publication Critical patent/WO2019105893A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54353—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals with ligand attached to the carrier via a chemical coupling agent
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells

Definitions

  • the present invention relates to a device for examining a cell suspension and a guided by using such a device by a method.
  • An example of such a method is flow cytometry, a technique in which cells are singulated and passed through a zone of defined electrical voltage or defined illumination at high speed. Depending on the shape, structure and / or coloration of the cells, different effects can be generated, on the basis of which certain properties of the cells can be derived. Downstream of an appropriate excitation and detection, the examined cells can, for example, be counted or optionally be diverted into different fluid channels and fed to specific collecting vessels.
  • an analysis of cell types, pathogens, proteins and the like on the basis of immunohistochemical properties is desirable.
  • immunoassays for example in anti- body-based detection methods such as ELISA
  • immobilize antibodies on a surface and to bind certain substances to them in this way, for example, proteins can be detected in blood.
  • laser microdissection An investigation by means of laser microdissection is also known in principle.
  • thin sections of tissue are applied to slides and can be stained there in a desired manner, in particular also using antibody-coupled dyes.
  • Correspondingly marked areas can be cut out in laser microdissection by means of a laser beam and finally transferred to a sample collecting vessel so that they can be accessed for subsequent analysis and diagnosis.
  • a disadvantage of known immunohistochemical assay methods such as ELISA is that there is generally no possibility of selectively releasing the binding partners which are bound to the used antibodies fixed on a surface, for example after a visual inspection or photometric evaluation, and further, eg molecular biological, to undergo analytical procedures. In such methods, either all binding partners or none at all can be extracted. Selection of specific binding partners is not possible.
  • a disadvantage of the use of laser microdissection methods is that their implementation typically requires complex sample preparation, in particular of cell suspensions, and the devices used for laser microdissection methods are complicated and expensive and sometimes demanding in operation. The same applies to flow cytometry.
  • the object of the present invention is to carry out improved possibilities for the investigation of cell suspensions, in particular on immunohistochemical bases.
  • the present invention is based, inter alia, on the finding that, in particular in microfluidic devices composed of supply, discharge and sorting lines, in particular in the form of capillaries, and examination chambers, there is a particularly advantageous investigation of cell suspensions immunohistochemical basis is possible.
  • An essential aspect of the present invention consists in equipping at least one chamber with optionally different binding partners for one or more different cell types.
  • the binding partners also referred to below as “affinity molecules”
  • the binding partners are anchored or immobilized on a surface of a corresponding chamber, which is also referred to below as an "interaction surface”, by means of a coupling molecule (linker) which is in particular photoactivatable.
  • the present invention thus operates with binding partners immobilized on a surface and not, for example, such as certain methods of flow cytometry, in particular FACS (fluorescence-activated cell sorting), with binding partners which are present in a liquid freely.
  • FACS fluorescence-activated cell sorting
  • cells are now flushed through the supply line into a corresponding examination chamber, some of these cells can bind to specific binding partners, depending on the expression of surface molecules or receptors (for example antigens), while other cells, if appropriate after a sufficient incubation period for binding, together with, for example, supernumerable cells capable of binding, can be flushed out of the chamber through the discharge line.
  • the bound cells can be visualized for example by means of a microscope system.
  • cells can either selectively from the bond with the Binding partner dissolved or selectively removed together with the corresponding binding partner of the interaction surface and transferred, for example by means of a sorting line in a specific analysis or further processing chamber or a corresponding receptacle. In this way, these cells are accessible for selective subsequent analysis or diagnosis.
  • the present invention proposes a device for assaying a cell suspension containing cells of a first and a second cell type.
  • different cell types may in particular be cells which, for example, present specific binding partners for specific affinity molecules on their surface. These may, for example, be antigens or other markers which are suitable for entering into a preferably specific binding with a particular affinity molecule, for example a corresponding antibody.
  • an interaction surface is used on which affinity molecules of a first and, in a further embodiment, of a second molecule type are fixed.
  • affinity molecules may be different antibodies that are capable of undergoing a known antigen-antibody binding or antigen-antibody reaction with the antigens presented on the different cell types.
  • antigen-antibody binding typically takes place according to the key-lock principle and can be extremely specific. An antigen-antibody binding can therefore make a particularly specific analysis possible in the context of the present invention.
  • affinity molecules include not only antibodies but also other molecules which enable a particularly selective binding to particular cell types. These may also be, for example, cell receptors or their ligands or certain cell-cell connection molecules such as, for example, desmosomes, cadherins, connexins or integrins, the last-mentioned affinity molecules connecting cells with the extracellular matrix in the biological sense.
  • cell receptors or their ligands or certain cell-cell connection molecules such as, for example, desmosomes, cadherins, connexins or integrins, the last-mentioned affinity molecules connecting cells with the extracellular matrix in the biological sense.
  • cell receptors or their ligands
  • certain cell-cell connection molecules such as, for example, desmosomes, cadherins, connexins or integrins, the last-mentioned affinity molecules connecting cells with the extracellular matrix in the biological sense.
  • a bond between the affinity molecules and their respective binding partners of the respective cell types is non-covalent
  • an affinity molecule comprises a radiation-activatable, for example photoactivatable, coupling molecule or a corresponding molecular segment (linker) via which it can be coupled, for example, to the interaction surface or can be coupled to the cells of the respective cell type.
  • a corresponding coupling molecule can also have a corresponding photoactivatable functional group whose excitation with suitable radiation leads to a detachment from the interaction surface or the respective cells.
  • Corresponding photoactivatable functional groups or their use for the reversible attachment of cells to surfaces are or are generally known from the specialist literature (see, for example, Kadern, LF et al., Rapid Reversible Photoswitching of Integrin-Mediated Adhesion at the Single Cell Level Adv.
  • Radical starters are molecules that are particularly easy to crack into free radicals.
  • photoactivatable radical starters can be used for the present invention.
  • free radical initiators are azobisisobutyronitrile, dibenzoyl peroxide, dilauroyl peroxide, di-tert-butyl peroxide, diisopropyl peroxydicarbonate and potassium peroxodisulfate.
  • the affinity molecules of the first molecule type have a greater binding affinity to the cells of the first cell type than to the cells of the second cell type
  • the affinity molecules of the second molecule type if present, have a greater binding affinity to the cells of the second cell type than to the cells of the first cell type.
  • the term binding affinity refers to, for example, in the case of antibodies, the tendency of a corresponding antibody to enter into antigen-antibody binding with a specific antigen. More generally, the binding affinity refers to a binding tendency expressed, for example, in the form of an affinity constant.
  • the affinity molecules of the first molecule type have no or a markedly low binding affinity to the cells of the second cell type.
  • the affinity molecules of the first molecule type are configured such that they exhibit radiation of a first radiation property, as explained below, for example, light of a defined first wavelength, of the interaction surface and / or a bond the cells of the first cell type are solvable.
  • the affinity molecules of the second molecule type if present, are designed in such a way that they are irradiated with radiation of the first or a second radiation property, in the case of the second radiation property, for example, light of a defined second wavelength, of the interaction surface and / or of a Binding with the cells of the second cell type are soluble.
  • the affinity molecules of the first molecule type and the affinity molecules of the second molecule type are present, and these are both solvable by the irradiation with the radiation of the first wavelength property from the interaction surface and / or the binding with the cells of the respective cell type
  • a targeted spatial irradiation can be carried out, for example, using point scanners or laser microdissection systems or setting a minimum possible aperture of a Microscope system can be done.
  • the cells or cells to be detached or to be detached in particular areas For example, types can be detected visually or automatically by evaluating a microscopic image.
  • the affinity molecules of the first molecule type and the affinity molecules of the second molecule type are present, and the affinity molecules of the first molecule type are irradiated by the irradiation with the radiation of the first radiation property, in particular selectively, from the interaction surface and / or the binding with the cells of the first cell type, however, the affinity molecules of the second molecule type, in particular selectively, by the irradiation with the radiation second radiation property of the interaction surface and / or the binding with the cells of the second cell type, is not necessarily one spatially resolved radiation is required.
  • a "selective" solubility or detachability is present when the affinity molecules of the first molecule type, but not the affinity molecules of the second molecule type can be solved by the irradiation with the radiation of the first radiation property or vice versa. It is also possible to use selectively and nonselectively removable affinity molecules. Of course, however, a spatially selective detachment can also be made in this case. In other words, in both cases an irradiation device can be provided or used which irradiates one or more selected regions of the interaction surface at least with the radiation of the first radiation property.
  • a corresponding cell type together with its respective affinity molecule is released from the interaction surface by the action of the radiation of the respective radiation property, but it can also be provided that the affinity molecules are formed in this way are that a bond between affinity molecule and the respective cell type is canceled by the radiation with the respective radiation property.
  • the action of the radiation with the defined radiation property removes the respective cell type together with the associated affinity molecule from the interaction surface and can thus be obtained
  • only the respective cell types are separated from their affinity molecules.
  • subsequent analysis may involve solving a bond between the affinity molecules and their respective cell types so that corresponding cells can be recovered individually.
  • an affinity molecule which remains bound to the interaction surface after exposure to radiation of the corresponding radiation type, may be available for interaction with further cells of the corresponding cell type so that a corresponding device is not "consumed".
  • the corresponding radiation responsiveness of the affinity molecules can be brought about in particular by the use of radiation-activatable, for example photoactivatable, coupling molecules or linkers, or of correspondingly excitable chemical groups.
  • radiation-activatable for example photoactivatable, coupling molecules or linkers
  • free-radical initiators of the type described above can be used as radiation- or photoactivatable coupling molecules.
  • the photoactivation of corresponding molecules or groups comprises at molecular level in particular the formation of free radicals by irradiation.
  • molecules can be used whose conformation can be changed by irradiation (for example from a cis to a trans conformation) and which change their binding properties as a result of this conformational change.
  • the affinity molecules of the first type of molecule may be, in particular, selective antibodies to antigens of the cells of the first cell type and the affinity molecules of the second molecule type, if present, to be selective antibodies to antigens of the second cell type.
  • mono- or polyclonal antibodies can be used, as is generally known from the prior art.
  • monoclonal antibodies play a role in diagnostics and research, and thus also in the context of the present invention, since they can bind a number of corresponding antigens with high specificity.
  • the respective named radiation is electromagnetic radiation, in particular light of a defined wavelength, in particular in the ultraviolet, wherein the radiation properties represent one or more specific wavelengths or wavelength ranges.
  • the first radiation property may have a first wavelength range and the second radiation characteristic.
  • genschaft if used, represent a second, different and in particular not overlapping with the first wavelength range wavelength range.
  • a corresponding "wavelength range” may in this case be a single wavelength, but typically corresponding photoactivatable coupling molecules respond to a certain bandwidth of different wavelengths, which is therefore selected accordingly and used for the detection.
  • corresponding narrowband or broadband light sources or corresponding filters can be used. These are preferably already integrated in a microscope system used for the examination.
  • a "radiation property" can also represent an intensity of the respectively used radiation, for example a specific light intensity. This is selected according to the respective coupling molecules to be examined or their specific light sensitivity.
  • the present invention is not limited to affinity molecules of two types of molecules or the investigation of cells of first and second cell types.
  • affinity molecules of a further molecule type can also be fixed on the interaction surface, the affinity molecules of the further molecule type then having a greater binding affinity to cells of a further cell type than to the cells of the first cell type and of the cells of the second cell type, and the affinity molecules of the further molecule type can be solubilized by irradiation with radiation of the first, the second or a further radiation property, in particular selectively, from the interaction surface and / or a bond with the cells of the further cell type.
  • affinity molecules of a further molecule type can also be fixed on the interaction surface, the affinity molecules of the further molecule type then having a greater binding affinity to cells of a further cell type than to the cells of the first cell type and of the cells of the second cell type, and the affinity molecules of the further molecule type can be solubilized by irradiation with radiation of the first, the second or a further radiation property, in particular selective
  • the device proposed according to the invention has a test chamber in which the interaction surface is arranged.
  • a corresponding examination chamber can have a defined volume into which a certain amount of cell suspension can be introduced for examination within the scope of the present invention.
  • the interaction surface forms a wall, for example a bottom surface, of a corresponding examination chamber and has a defined size.
  • an interaction surface in - Kl a corresponding chamber (when in use) be arranged horizontally, so that it overlays when using the device by means of a cell suspension and in particular visually from top or bottom, for example, macroscopic or microscopic, can be inspected.
  • the examination chamber is set up for a microscopic inspection of the interaction area.
  • the examination chamber may be designed to be transparent on a side facing away from the interaction surface, for example by using glass or a transparent plastic.
  • the device is designed in particular as a microfluidic device, so that a side of the examination chamber facing away from the interaction surface can be designed, for example, in the manner of a conventional cover glass with a thickness of 0.17 mm and can be arranged close above the interaction surface.
  • the interaction surface itself in a corresponding examination chamber in a transparent manner and in a suitable thickness, so that in this way a visual microscopic inspection of the interaction surface or a recording of a corresponding especially digital, image is possible.
  • This can be done for example by means of an inverted microscope system, in which an observation beam path is directed from below onto a corresponding interaction surface.
  • use in a regular, i. not inverse microscope system in which the interaction surface, if it is transparent and designed for examination can be arranged on an upper side of an examination chamber, which in this case can be flooded with the cell suspension.
  • Other configurations of corresponding examination chambers for a microscopic inspection are possible in principle.
  • a microfluidic device in which the examination chamber is in particular connected to a first line, with which the cell suspension can be fed into the examination chamber.
  • a first line may in particular be designed as a capillary, which may be provided at an end facing away from the examination chamber, with an inlet opening, which may be provided, for example, with a pump. Pettenspitze is coupled and thus allows a particularly advantageous filling.
  • a corresponding line can also be designed to be closable, so that, for example, evaporation of liquid from the examination chamber, which, for example, is also spent for a specific incubation period in an incubator and can be heated there, for example, can be prevented.
  • the examination chamber in the device according to the invention is further connected to a second line, by means of which fluid can be withdrawn from the examination chamber.
  • a second line by means of which fluid can be withdrawn from the examination chamber.
  • a corresponding examination chamber can also be rinsed in particular via the first and the second line, so that excess material can be easily transported away and thus, for example, cross contamination with undesired cell types can be prevented.
  • the second conduit can be fluidically connected optionally to a third and a fourth line.
  • the second line can therefore branch into two further lines into which, in particular, the cells of the respective cell types can be removed after a corresponding action of radiation. In this way, selective recovery of corresponding different cell types, for example in different collecting vessels, is possible. It can also be provided to connect the examination chamber directly to several lines. In this case, the second line is not required.
  • the present invention also extends to a method for assaying a cell suspension containing cells of a first and a second cell type, such a device using a device as has been explained above in different embodiments, ie a device with an interaction surface on which at least affinity molecules of a first molecule type are fixed, wherein the affinity molecules of the first molecule type have a greater binding affinity to the cells of the first cell type than to the cells of the second cell type, and wherein the affinity molecules of the first molecule type by irradiation with radiation a first radiation property from the interaction surface and / or from a binding Rank are soluble with the cells of the first cell type.
  • the method according to the invention therefore benefits from the advantages explained above in the same way, so that it can be expressly referred to.
  • the method according to the invention comprises immobilizing the cells of the first cell type at the interaction surface via the affinity molecules of the first molecule type by contacting the cell suspension with the interaction surface for an interaction period.
  • a corresponding interaction period can be selected according to the speed or reaction kinetics of a corresponding reaction.
  • a further step after such a contact may include, in particular, the emptying and / or rinsing of the interaction surface, so that only the particular cell types bound remain on the corresponding interaction surface.
  • the cells of the first cell type are released with the affinity molecules of the first molecule type or of the cells of the first cell type without the affinity molecules of the first molecule type from the interaction surface by irradiating the interaction surface or the affinity molecules fixed thereto with the radiation of the first radiation type.
  • irradiation can also take place for a given irradiation time or a first irradiation period.
  • a visual inspection of the interaction surface with the cells bound thereto in particular using a microscope, can take place.
  • the cells of the first cell type released by irradiation with the radiation of the first radiation type can be collected with the affinity molecules of the first molecule type or the released cells of the cell type without the affinity molecules of the first molecule type.
  • corresponding cells can also be transported selectively into a collecting vessel and, for example, counted.
  • affinity molecules of a second molecule type are fixed on the interaction surface in addition to the affinity molecules of the first molecule type, the affinity molecules of the second molecule type having a larger molecular weight Binding affinity to the cells of the second cell type than to the cells of the first cell type, and wherein the affinity molecules of the second molecule type are irradiated with the radiation of the first radiation property or by irradiation with radiation of a second radiation property from the interaction surface and / or from a bond with the cells of the second cell type are solvable.
  • immobilization of the cells of the second cell type at the interaction surface via the affinity molecules of the second molecule type takes place in particular by contacting the cell suspension with the interaction surface for the interaction period.
  • Release of the cells of the second cell type with the affinity molecules of the second molecule type or the cells of the second cell type without the affinity molecules of the second molecule type from the interaction surface according to this embodiment by irradiating the interaction surface with the radiation of the first radiation type or by irradiating the interaction surface with the radiation of the second radiation type, wherein, if applied, irradiation of the interaction surface with the radiation of the second radiation type is carried out in particular after the cells of the first cell type have been previously collected, ie in a further one (later) irradiation period.
  • one or more selected areas of the interaction surface can be irradiated. Advantages of such a procedure have already been explained.
  • each released cell may include flushing the released cells by means of a fluid over one or more conduits.
  • FIG. 1 shows a device according to an advantageous embodiment of the present invention in a simplified, schematic representation.
  • Figures 2A to 2E illustrate an examination of a cell suspension according to a particularly preferred embodiment of the present invention.
  • FIG. 3 illustrates a method according to a particularly preferred embodiment of the present invention.
  • a device according to a particularly preferred embodiment of the present invention is shown schematically in simplified form.
  • the device is designated overall by 100.
  • the device 100 is a microfluidic device, which may be designed, for example, in the form of a so-called "chip", in particular in the size of a conventional microscope slide or in a size order comparable thereto.
  • the device 100 comprises an examination chamber 20 which has an interaction surface which is still shown in detail in the following FIGS. 2A to 2E.
  • the examination chamber 20 is connected to a first line 24, by means of which a cell suspension can be fed into the examination chamber 20. It is furthermore connected via a second line 25 to a third line 26 and a fourth line 27.
  • the second line 25 By means of the second line 25, fluid can be withdrawn from the examination chamber 20 and optionally distributed to the third line 26 and to the fourth line 27. In this way, cells removed from the sample chamber 20 can be selectively counted and / or collected.
  • the third line 26 and the fourth line 27 corresponding lines can also be coupled directly to the examination chamber.
  • the second line 25 may also be omitted.
  • FIGS. 2A to 2E a study of a cell suspension according to a particularly preferred embodiment of the present invention is simplified in detail and schematically illustrated.
  • the function of the interaction surface 23 is illustrated in FIGS. 2A to 2E.
  • Affinity molecules 21, 22 of different types are illustrated on the interaction surface 23. These are symbolized as antibodies in the example shown. In the illustrated example, they are coupled to the interaction surface via photoactivatable linkers 21a, 22a. In this way, the interaction molecules 21, 22 can each be detached from the interaction surface by radiation of a specific radiation property, for example light of specific wavelengths. According to FIG. 2A, no cell suspension is introduced into a corresponding device.
  • the interaction surface 23 may be overlaid in this phase, for example, to protect it with a buffer of appropriate type.
  • FIG. 2B shows how cells of different cell types, designated here by 1 1 and 12, each attach to the affinity molecules 21, 22 of the different types of molecules or bind to them.
  • the interaction surface 23 is overlaid with a cell suspension containing cells 11, 12 of corresponding cell types.
  • the cell suspension can be removed from the interaction surface 23, for example by aspiration of excess cells that are not bound to the affinity molecules 21, 22.
  • a selective activation of the affinity molecules 21, here of the first type of molecule can now take place. This results in a detachment of the corresponding cells 11 with the respective affinity molecules 21 from the interaction surface 23, as illustrated in FIG. 2D, so that these cells 11 can be selectively obtained.
  • FIG. 2E illustrates an alternative of a corresponding activation, which involves a detachment of the respective cells 1 1 from their activity molecules 21, but the latter remain bound to the interaction surface 23.
  • an activation of the affinity molecules 22 and the detachment of the corresponding cells 12 can also take place. Appropriate steps can be continued as long as appropriate affinity molecules and their associated cell types are present.
  • FIG. 3 shows a method in accordance with a particularly preferred embodiment of the present invention in the form of a simplified, schematic flow chart, designated overall by 200.
  • the method 200 as illustrated here in a simplified manner, comprises three method steps 210, 220, and 230.
  • step 210 at least a part of a cell suspension is brought into contact with an interaction surface for an interaction period and thereby binding the cells of a first cell type to the affinity molecules of the first molecule type.
  • binding of the cells of the second cell type to the affinity molecules of the second molecular type may also take place here, if present.
  • the interaction surface is irradiated with the radiation of the first radiation type for a first irradiation period and thereby dissolving the cells of the first cell type with the affinity molecules of the first molecule type from the interaction surface and / or the cells of the first cell type the activity molecules of the first molecule type.
  • a subsequent step 230 collection of the cells of the first cell type released by irradiation with the radiation of the first radiation type with the affinity molecules of the first molecule type or the released cells of the first cell type can be carried out without the affinity molecules of the first molecule type.
  • method steps 210 to 230 further method steps may follow, in particular for obtaining further cell types by appropriate action of radiation, if further affinity molecules for corresponding further cell types are present.
  • a visual or microscopic inspection can also be provided in any of the method steps 210 to 230 shown.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Food Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention concerne un dispositif (100) d'examen d'une suspension de cellules (10) qui contient des cellules (11, 12) d'un premier et d'un deuxième type de cellules, le dispositif comportant une surface d'interaction (23) sur laquelle sont fixées des molécules d'affinité (21) d'un premier type de molécules, les molécules d'affinité (21) du premier type de molécules présentant une affinité de liaison aux cellules (11) du premier type de cellules supérieure à son affinité de liaison aux cellules (12) du deuxième type de cellules, et les molécules d'affinité (21) du premier type de molécules étant solubles pouvant être libérées par une irradiation avec un rayonnement d'une première propriété de rayonnement de la surface d'interaction (23) et/ou une liaison avec les cellules (11) du premier type de cellules. L'invention a également pour objet un procédé qui consiste en l'utilisation d'un dispositif correspondant.
PCT/EP2018/082576 2017-12-01 2018-11-26 Dispositif et procédé d'examen d'une suspension de cellules Ceased WO2019105893A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102017128627.6A DE102017128627B4 (de) 2017-12-01 2017-12-01 Vorrichtung und Verfahren zur Untersuchung einer Zellsuspension
DE102017128627.6 2017-12-01

Publications (1)

Publication Number Publication Date
WO2019105893A1 true WO2019105893A1 (fr) 2019-06-06

Family

ID=64564857

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2018/082576 Ceased WO2019105893A1 (fr) 2017-12-01 2018-11-26 Dispositif et procédé d'examen d'une suspension de cellules

Country Status (2)

Country Link
DE (1) DE102017128627B4 (fr)
WO (1) WO2019105893A1 (fr)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8906700B2 (en) * 2007-11-06 2014-12-09 Ambergen, Inc. Methods and compositions for phototransfer

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014141386A1 (fr) * 2013-03-12 2014-09-18 株式会社日立製作所 Dispositif à réseau cellulaire bidimensionnel et appareil pour la quantification de gènes et l'analyse de séquences

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8906700B2 (en) * 2007-11-06 2014-12-09 Ambergen, Inc. Methods and compositions for phototransfer

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
KADEM, L.F. ET AL.: "Rapid Reversible Photoswitching of Integrin-Mediated Adhesion at the Single-Cell Level", ADV. MATER., vol. 28, no. 9, 2016, pages 1799 - 1802
SHINYA ARIYASU ET AL: "Selective Capture and Collection of Live Target Cells Using a Photoreactive Silicon Wafer Device Modified with Antibodies via a Photocleavable Linker", LANGMUIR, vol. 28, no. 36, 11 September 2012 (2012-09-11), US, pages 13118 - 13126, XP055363606, ISSN: 0743-7463, DOI: 10.1021/la302393p *

Also Published As

Publication number Publication date
DE102017128627A1 (de) 2019-06-06
DE102017128627B4 (de) 2019-08-14

Similar Documents

Publication Publication Date Title
EP1311655B1 (fr) Dispositif et procede pour mettre en contact electrique des cellules biologiques en suspension dans un liquide
DE69222826T2 (de) Behälter für flüssige proben und befestigbare test-module
DE69804239T2 (de) Verfahren und apparat zur durchmischung und abscheidung von partikel-material aus einer flüssigen probe
DE69303483T2 (de) Mikrohergestellte Vorrichtungen zum Handhaben von Sperma
DE3586983T2 (de) Verfahren und vorrichtung fuer immunotest.
DE602005005485T2 (de) Assayvorrichtung und verfahren mit gesteuertem fluss
EP2152913B1 (fr) Dispositif de détection permettant la détection de microparticules biologiques telles que des bactéries, des virus, des spores, du pollen ou des toxines biologiques, et procédé de détection
DE60034743T2 (de) Verfahren und Vorrichtung zur Abtrennung von festförmigen Bestandteilen aus einem flüssigen Bestandteil in einem biologischen Flüssigkeit
EP3380825B1 (fr) Dispositif et procédé pour analyser des objets biologiques par spectroscopie raman
DE60107538T2 (de) Vorrichtung zum trennen und nachweis von bewegungsfähigen spermatozoen
EP1590652A1 (fr) Procede de triage et d'identification de cellules a plusieurs parametres et dispositif correspondant
DE102011076221B3 (de) Automatisierter CTC Nachweis
EP4326440A1 (fr) Procédé de détection de cellules contenant un noyau dans un échantillon liquide d'un patient au moyen d'un dispositif microfluidique et dispositif microfluidique
EP3039399A1 (fr) Système de microdissection par laser et procédé pour système de microdissection par laser
EP2142911A2 (fr) Biopuce pour l'analyse par fluorescence de transporteurs individuels
DE10148210B4 (de) Flusskammer
DE3883652T2 (de) Verfahren zum sortieren von zellen.
DE102017128627B4 (de) Vorrichtung und Verfahren zur Untersuchung einer Zellsuspension
DE60031521T2 (de) Produkte und verfahren für einzelwert- und vielfachwert-phänotypisierung von zellen
DE102020107645B4 (de) Betrieb einer Mikrofluidik-Vorrichtung bei der Analyse von Probesubstanzen
DE602004007429T2 (de) Verfahren zur Zellselektion
DE102008056583A1 (de) Verfahren und Vorrichtung zur Feststellung der Reagenzienqualität
DE202013102057U1 (de) Vorrichtung zur Extraktion von Nukleinsäuren mittels magnetischer Beads
DE202007014762U1 (de) Anordnung zur Aufbereitung von zu untersuchendem Gewebe sowie Objektträger für zu untersuchendes Gewebe
EP1367380A1 (fr) Méthode et dispositif pour la localisation et manipulation automatique de cellules

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18811772

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18811772

Country of ref document: EP

Kind code of ref document: A1